CN109932345A - A kind of lysine detection method based on quantum dot and nanogold - Google Patents
A kind of lysine detection method based on quantum dot and nanogold Download PDFInfo
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Abstract
The present invention provides a kind of based on the highly selective method with Sensitive Detection lysine of quantum dot and nanogold reversible fluorescence, the fluorescence intensity change of cadmium quantum dot and modified Nano gold mixed system before and after various concentration lysine is added by measurement, and its linear relationship with lysine concentration is established, realization mainly includes fluorescence quantum to the content detection of lysine.For the fluorescence quantum that the present invention uses for cadmium telluride quantum dot, nanogold is the nanogold of dodecyl trimethyl ammonium bromide self assembly citrate sealing end, can form a kind of new reversible fluorescence probe；The reversible fluorescence probe can efficiently, quickly and accurately detect the content of lysine, and detection range is 0.5-100 μm of ol/L, and detection limit can reach 3.22nmol/L；And the synthesis process being related to is simple, analysis cost is cheap, and is remarkably improved the selectivity to lysine detection.
Description
Technical field
The invention belongs to technical field of analysis and detection, and in particular to a kind of to be detected based on quantum dot and the lysine of nanogold
Method.
Background technique
Lysine is one of eight big essential amino acids of human body, is played a crucial role to human body metabolism's process,
Important intermediate of the lysine as metabolic pathway can promote human development, enhancing immune function, and can play raising maincenter mind
Effect through function of organization.But lysine metabolism will lead to certain physiological function diseases extremely, such as high sarcosine disease,
Blood or cystinuria etc..
Currently, having had attempted to develop numerous sensors for being used to detect lysine, it is such as based on carbon dots, nanogold, sulphur
The sensing system of element-T- heparin etc., it is right simultaneously although these sensors can fast implement the analysis detection of lysine
Arginine, histidine etc. have a degree of response, to interfere the accurate quantitative analysis of lysine.In addition conventional method is to lysine
Response sensitivity it is poor, therefore the sensor of interest for further developing a kind of high sensitivity be still lysine detection technique urgently
A big problem to be solved.
Summary of the invention
It is a primary object of the present invention to ask for existing lysine detection technique poor specificity, detection process are cumbersome etc.
Topic, provides a kind of lysine detection method based on quantum dot and nanogold, Gao Te of this method based on nanogold and lysine
Opposite sex reaction, and using quantum dot as fluorescence sense, nano material is combined with sensing technology and constructs a kind of reversible fluorescence
Sensor can show excellent selectivity and sensitivity to lysine, and the detection method being related to is easy, and suitable popularization is answered
With.
To achieve the above object, the technical solution adopted by the present invention are as follows:
A kind of lysine detection method based on quantum dot and nanogold, includes the following steps:
1) fluorescence intensity of quantum dot and modified Nano gold mixed system before and after various concentration lysine is added is measured respectively
F0And F1, and calculate fluorescence intensity change (F1-F0)/F0；
2) fluorescence intensity change (F is established1-F0)/F0With the linear relationship of lysine concentration, the content to lysine is realized
Detection.
In above scheme, the cadmium telluride quantum dot is CdTe quantum, and preparation method includes the following steps: to take cadmium salt
It is dispersed in water, stirs evenly with N-acetyl-L-cysteine, and the pH value for adjusting acquired solution system seals ice bath to 8
Afterwards, it is filled with nitrogen thereto and stirs；Then plus tellurium source continues to stir, and is finally transferred in reaction kettle and heats instead at 200 DEG C
Answer 1 hour aqueous solution to get fluorescence quantum CdTe.
In above scheme, the modified Nano gold is to be sealed using the self-assembled modified citrate of cationic surfactant
The nanogold at end.
In above scheme, dodecyl trimethyl ammonium bromide or cetyl three is can be selected in the cationic surfactant
Methyl bromide ammonium etc..
In above scheme, the preparation method of the modified Nano gold includes the following steps:
I) citrate is dispersed in water, gold chloride is added under agitation and boiling water, microwave heating are reacted,
The nanogold of cooling citrate sealing end；
Ii) nanogold that citrate blocks is put and is added in cationic surfactant solution, at a temperature of 37-60 DEG C
Hatch 30-40min, it is cooling to get the modified Nano gold.
In above scheme, the molar ratio of the citrate and gold chloride is 1:(0.3~0.7).
In above scheme, microwave heating power described in step i) is 750~950W, and the time is 3~5min.
In above scheme, the nanogold of the citrate sealing end and the mass ratio of cationic surfactant are 1:
(1.20×103~1.86 × 103)。
In above scheme, in the quantum dot and modified Nano gold mixed system, the concentration of quantum dot is 4.6 × 10-9~
7.2×10-8mol/L；The concentration of modified Nano gold is 3 × 10-9~5.0 × 10-7mol/L。
In above scheme, the fluorescence intensity is the fluorescence intensity under 360nm excitation wavelength at 630nm.
The principle of the present invention are as follows:
The present invention is by the fluorescence between the modified Nano gold and cadmium telluride quantum dot of cationic surfactant self assembly
Resonance energy transfer (the ultra-violet absorption spectrum red shift of promotion nanogold is simultaneously Chong Die with the emission spectrum of quantum dot) and electrostatic interaction
Cadmium telluride quantum dot fluorescence is caused to quench, and by lysine and the modified Nano of cationic surfactant self assembly gold
With the specific reaction of lysine (unmodified nanogold to lysine without response, and DTAB lysine is responded it is very weak), make
The fluorescence of cadmium telluride quantum dot releases, and forms a kind of reversible fluorescence probe, and quickly embodies the fluorescence after introducing lysine
Strength Changes (at 630nm), and the fluorescence intensity change and lysine concentration are in a linear relationship, can pass through cadmium telluride quantum dot-
The concentration of nanogold reversible fluorescence probe in detecting lysine.
Compared with prior art, the invention has the benefit that
1) detection method that invention is related to is simple, and can realize the detection of lysine concentration with high specificity, can be bad ammonia
The efficient detection technology of acid provides a new thought.
2) selectivity and high sensitivity that detection method of the present invention detects lysine concentration are, it can be achieved that 0-100 μ
To the efficient detection of lysine within the scope of mol/L, it is effectively improved the problems such as conventional method is poor to the response sensitivity of lysine.
3) synthesis technology of the present invention is simple, reaction condition is mild, at low cost, is suitble to promote and apply.
Detailed description of the invention
Fig. 1 is the fluorescence intensity curves of the different mixed systems of 1 gained of embodiment；
Fig. 2 is fluorescence response figure of the different aminoacids described in embodiment 2 to reversible fluorescence probe；
Fig. 3 is 3 gained cadmium quantum dot of embodiment and modified Nano gold mixed system to lysine (2,6- diamino is sour)
A, the fluorescence intensity curves of 1,5- diaminovaleric acid B, 6-aminocaprolc acid C and Amicar D；
Fig. 4 is the fluorescence that various concentration lysine is added to cadmium quantum dot and modified Nano gold mixed system in embodiment 4
Intensity variation curve；
Fig. 5 is the linear relationship of fluorescence intensity change (F0-F1)/F0 and lysine concentration.
Fig. 6 is fluorescence intensity change curve described in comparative example 1.
Fig. 7 is figure of fluorescence intensity changes described in comparative example 1.
Specific embodiment
For a better understanding of the present invention, below with reference to the embodiment content that the present invention is furture elucidated, but it is of the invention
Content is not limited solely to the following examples.
In following embodiment, for CdTe quantum, preparation method includes the following steps: to take 0.12g the quantum dot used
Cadmium Chloride powder and 0.09g N-acetyl-L-cysteine powder are scattered in 40mL secondary distilled water, are stirred 15min, are then used
After sealing ice bath, 20min is stirred when being filled with nitrogen thereto to 8 for sodium hydrate regulator solution pH value；It is subsequently added into 0.02g
Sodium tellurite continues to stir, and is finally transferred in reaction kettle heating reaction 1 hour at 200 DEG C, can be obtained CdTe fluorescence volume
The aqueous solution of son point.
In following embodiment, the modified Nano gold used the citric acid self-assembled modified for dodecyl trimethyl ammonium bromide
The nanogold of salt sealing end, preparation method include the following steps:
1) 0.29g monohydrate potassium sodium is taken to be scattered in 1mL secondary distilled water, 2mL concentration is added while stirring is
The chlorauric acid solution and 98mL boiling water (secondary distilled water) of 0.3mol/L, moves in micro-wave oven and heats 3min under 950W power,
It is cooled to room temperature, can be obtained the nanogold of citrate sealing end；
2) nanogold and cationic surfactant for taking the sealing end of citrate obtained by step 1) are by 1:1.52 × 103Matter
Amount is cooled to room temperature after being then incubated for half an hour at 37 DEG C, can be obtained receiving for cationic surfactant self assembly than mixing
Meter Jin (modified Nano gold).
Embodiment 1
A kind of lysine detection method (feasibility study) based on quantum dot and nanogold, includes the following steps:
1) the 100 μ L of cadmium telluride quantum dot solution of 552nmol/L is taken, 900 μ L secondary distilled waters are added in 360nm excitation wave
Its fluorescence intensity at 630nm is measured under length, is recorded as F (the result is shown in Figure 1 A)；
2) take 100 μ L of cadmium telluride quantum dot solution of 552nmol/L, 90 μ L of modified Nano gold solution of 47.2nmol/L,
810 μ L secondary distilled waters, its fluorescence intensity at 630nm is measured under 360nm excitation wavelength, is recorded as F after mixing0(knot
Fruit sees Fig. 1 C)；
3) take 100 μ L of cadmium telluride quantum dot solution of 552nmol/L, 90 μ L of modified Nano gold solution of 47.2nmol/L,
The 100 μ L of lysine solution and 710 μ L secondary distilled waters of 1mmol/L, measured under 360nm excitation wavelength after mixing its
Fluorescence intensity under 630nm, is recorded as F1(the result is shown in Figure 1 B).
As seen from Figure 1, the modified Nano gold and cadmium telluride amount of present invention gained cationic surfactant self assembly
Fluorescence resonance energy transfer and electrostatic interaction between son point cause cadmium telluride quantum dot fluorescence to quench；After introducing lysine
Specific reaction will occur for the modified Nano gold and lysine of cationic surfactant self assembly, make the glimmering of cadmium telluride quantum dot
Light releases, and can form a kind of reversible fluorescence probe, realizes quick, efficient detection to lysine.
Embodiment 2
A kind of lysine detection method (highly selective research) based on quantum dot and nanogold, includes the following steps:
1) take 100 μ L of cadmium telluride quantum dot solution of 552nmol/L, 90 μ L of modified Nano gold solution of 47.2nmol/L,
810 μ L secondary distilled waters, its fluorescence intensity at 630nm is measured under 360nm excitation wavelength, is recorded as F after mixing0；
2) take 100 μ L of cadmium telluride quantum dot solution of 552nmol/L, 90 μ L of modified Nano gold solution of 47.2nmol/L,
100 13 kinds of μ L amino acid solutions (100 μm of ol/L of lysine；Serine, histidine, leucine, proline, serine, asparagus fern
Propylhomoserin, threonine, arginine, phenylalanine, glutamic acid, isoleucine, methionine concentration are 1000 μm of ol/L), 710 μ L bis-
Secondary distilled water measures its fluorescence intensity at 630nm under 360nm excitation wavelength, is recorded as F after mixing1；
3) fluorescence intensity change (F after different aminoacids solution is added is calculated1-F0)/F0；As a result see Fig. 2.
As seen from Figure 2, the reversible fluorescence probe based on quantum dot and nanogold obtained by the present embodiment has lysine
Have highly selective.
Embodiment 3
A kind of lysine detection method (special Journal of Sex Research) based on quantum dot and nanogold, includes the following steps:
1) take 100 μ L of cadmium telluride quantum dot solution of 552nmol/L, 90 μ L of modified Nano gold solution of 47.2nmol/L,
810 μ L secondary distilled waters, its fluorescence intensity at 630nm is measured under 360nm excitation wavelength, is recorded as F after mixing0；
2) take 100 μ L of cadmium telluride quantum dot solution of 552nmol/L, 90 μ L of modified Nano gold solution of 47.2nmol/L,
100 μ L lysine solutions (2,6- diamino sour) A and lack carboxyl, α-NH compared to lysine2With ε-NH21,5- diamino
Base valeric acid solution (B), 6-aminocaprolc acid solution (C) and Amicar solution (D) add 710 μ L secondary distilled waters, mix
Its fluorescence intensity at 630nm is measured after conjunction under 360nm excitation wavelength, is recorded as F1；
3) fluorescence intensity change (F after different aminoacids solution is added is calculated1-F0)/F0；As a result see Fig. 3.
As seen from Figure 3, only lysine is to cadmium telluride quantum dot and dodecyl trimethyl ammonium bromide self assembly lemon
The stable nanogold of lemon hydrochlorate has fluorescence recovery effects, shows that the special amino of lysine and carboxyl can be with the reversible fluorescences
Probe specificity effect.
Embodiment 4
A kind of lysine detection method (Code's Sensitivity) based on quantum dot and nanogold, includes the following steps:
1) take 100 μ L of cadmium telluride quantum dot solution of 552nmol/L, 90 μ L of modified Nano gold solution of 47.2nmol/L,
810 μ L secondary distilled waters, its fluorescence intensity at 630nm is measured under 360nm excitation wavelength, is recorded as F after mixing0；
2) take cadmium telluride quantum dot solution 100 the μ L, 47.2nmol/L of 552nmol/L 90 μ L of modified Nano gold solution,
100 μ L various concentration lysines (0-100 μm of ol/L), add 710 μ L secondary distilled waters, in 360nm excitation wavelength after mixing
Lower its fluorescence intensity at 630nm of measurement, is recorded as F1, fluorescence intensity change curve is shown in Fig. 4；
3) fluorescence intensity change (F after various concentration lysine solution is added is calculated1-F0)/F0；Gained fluorescence intensity becomes
Change (F0-F1)/F0Fig. 5 is seen with the linear relationship of lysine concentration.
As seen from Figure 4, increase with lysine concentration, fluorescence intensity enhancing of the reversible fluorescence probe at 630nm；And
Fluorescence intensity change (F0-F1)/F0It is in a linear relationship with lysine concentration, it may be implemented within the scope of 0-100 μm of ol/L to lysine
Detection.
Comparative example 1
A kind of lysine detection method based on quantum dot and nanogold, includes the following steps:
1) the 100 μ L of cadmium telluride quantum dot solution of 552nmol/L is taken to be added into 900 μ L secondary distilled waters, after mixing in
Its fluorescence intensity at 630nm is measured under 360nm excitation wavelength, is recorded as F0；
2) take the 100 μ L of cadmium telluride quantum dot solution of three parts of 552nmol/L and the mixing of 810 μ L secondary distilled waters molten respectively
Liquid is separately added into nanogold (AuNPs) solution of the 47.2nmol/L of 90 μ L, the modified Nano gold of 47.2nmol/L thereto
(DTAB-AuNPs) solution and 1.2 × 10-3The DTAB solution of mol/L mixes, and then measures it respectively under 360nm excitation wavelength
Fluorescence intensity at 630nm, is recorded as F1, F2, F3(fluorescence intensity change curve is shown in Fig. 6)；
As seen from Figure 6, with the addition of AuNPs, the fluorescence intensity F of cadmium telluride quantum dot1It does not quench not only, very
To faint humidification is generated, show that unmodified nanogold can not quench the fluorescence of cadmium telluride quantum dot.And it is being added
The fluorescence intensity of cadmium telluride quantum dot weakens after DTAB and DTAB-AuNPs, but modified nanogold F2(DTAB-AuNPs) sudden
Effect of going out is more significant, shows that modified nanogold plays a crucial role the quenching of quantum dot.
Comparative example 2
A kind of lysine detection method (comparative experimental study) based on quantum dot and nanogold, includes the following steps:
1) the 100 μ L of cadmium telluride quantum dot solution of 552nmol/L is taken to be added into 900 μ L secondary distilled waters, after mixing in
Its fluorescence intensity at 630nm is measured under 360nm excitation wavelength, is recorded as F0
2) take the 100 μ L of cadmium telluride quantum dot solution of three parts of 552nmol/L and the mixing of 810 μ L secondary distilled waters molten respectively
Liquid, being separately added into 90 μ L concentration thereto is nanogold (AuNPs) solution of 47.2nmol/L, 1.2 × 10-3The DTAB of mol/L
Solution, 4.72nmol/L golden (DTAB-AuNPs) the solution mixing of modified Nano after measure it respectively under 360nm excitation wavelength
Fluorescence intensity at 630nm, is recorded as F1, F2, F3。
3) the 100 μ L of cadmium telluride quantum dot solution of three parts of 552nmol/L is taken to mix with 710 μ L secondary distilled waters respectively again molten
Liquid, being separately added into 90 μ L concentration thereto is nanogold (AuNPs) solution of 47.2nmol/L, 1.2 × 10-3The DTAB of mol/L
Golden (DTAB-AuNPs) solution of modified Nano of solution, 4.72nmol/L, then adding 100 μ L concentration respectively is 100 μm of ol/
The lysine solution of L measures its fluorescence intensity at 630nm respectively under 360nm excitation wavelength and is recorded as F after mixing1',
F2', F3' (fluorescence intensity change is shown in Fig. 7)；
As seen from Figure 7, modified nanogold plays a crucial role the quenching of quantum dot；And lysine
Addition the fluorescence intensity after DTAB and DTAB-AuNPs quenching CdTe quantum can be enhanced, but lysine is to DTAB-AuNPs
Recovery effects (the F of fluorescence intensity after quenching quantum dot3') become apparent, this is attributed between lysine and modified Nano gold
Specific effect.
Each raw material cited by the present invention can realize that the bound value of the present invention and each raw material, interval value can
Realize the present invention；Embodiment numerous to list herein.The bound value of technological parameter of the invention, interval value can realize this
Invention, embodiment numerous to list herein.
Claims (10)
1. a kind of lysine detection method based on quantum dot and nanogold, includes the following steps:
1) the fluorescence intensity F of the quantum dot and modified Nano gold mixed system before and after various concentration lysine is added is measured respectively0With
F1, and calculate fluorescence intensity change (F1-F0)/F0；
2) fluorescence intensity change (F is established1-F0)/F0With the linear relationship of lysine concentration, the content detection to lysine is realized.
2. lysine detection method according to claim 1, which is characterized in that the quantum dot is CdTe quantum.
3. lysine detection method according to claim 1, which is characterized in that the modified Nano gold is to utilize cation
Surfactant carries out the nanogold of self-assembled modified citrate sealing end.
4. lysine detection method according to claim 3, which is characterized in that the cationic surfactant is 12
Alkyl trimethyl ammonium bromide or cetyl trimethylammonium bromide.
5. lysine detection method according to claim 3, which is characterized in that the preparation method packet of the modified Nano gold
Include following steps:
I) citrate is dispersed in water, gold chloride is added under agitation and boiling water, microwave heating are reacted, it is cooling
Obtain the nanogold of citrate sealing end；
Ii) nanogold that citrate blocks is put and is added in cationic surfactant solution, is hatched at a temperature of 37-60 DEG C
30-40min, it is cooling to get the modified Nano gold.
6. lysine detection method according to claim 5, which is characterized in that mole of the citrate and gold chloride
Than for 1:(0.3~0.7).
7. lysine detection method according to claim 5, which is characterized in that microwave heating power described in step i) is
750~950W, time are 3~5min.
8. lysine detection method according to claim 5, which is characterized in that the nanogold of citrate sealing end with
The mass ratio of cationic surfactant is 1:(1.20 × 103~1.86 × 103)。
9. lysine detection method according to claim 1, which is characterized in that the quantum dot and the mixing of modified Nano gold
In system, the concentration of quantum dot is 4.6 × 10-9~7.2 × 10-8mol/L；The concentration of modified Nano gold is 3 × 10-9~5.0 ×
10-7mol/L。
10. lysine detection method according to claim 1, which is characterized in that the fluorescence intensity is 360nm excitation wave
Fluorescence intensity at long lower 630nm.
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